PT1988786E - Oligosaccharide mixture - Google Patents

Abstract

An oligosaccharide mixture comprising 5-70 wt % of at least one N-acetylated oligosaccharide selected from the group comprising GalN Acα1,3Galβ1,4Glc and Galβ1,6GalN Acα1,3Galβ1,4Glc, 20-90 wt % of at least one neutral oligosaccharide selected from the group comprising Galβ1,6Gal, Galβ1,6Glβ1,4Glc Galβ1,6Galβ1,6Glc, Galβ1,Galβ1,3Glc, Galβ1,3Galβ1,4Glc, Galβ1,6Galβ1,6Galβ1,4Glc, Galβ1,6Galβ1,3Galβ1,4Glc, Galβ1,3Galβ1,6Galβ1,4Glc and Galβ1,3Galβ1,3Galβ1,4Glc and 5-50 wt % of at least one sialylated oligosaccharide selected from the group comprising NeuAcα2,3Galβ1,4Glc and NeuAcα2,6Galβ1,4Glc; food products comprising said oligosaccharide mixture.

Description

DESCRIPTION " MIXTURE OF OLIGOSACCHARIDES "

Field of the Invention The invention relates to a mixture of oligosaccharides, to food products comprising said oligosaccharide mixture and to processes for the production of said oligosaccharide mixture.

BACKGROUND OF THE INVENTION The human colon is colonized with a broad range of bacteria that have positive and negative effects on bowel physiology, in addition to having other systemic influences. The groups of predominant bacteria found in the colon include bacteroids, bifidobacteria, eubacteria, clostridias and lactobacilli. The bacteria present have buoyant activities in response to the availability of substrates, redox potential, pH, O 2 tension and distribution in the colon. In general, intestinal bacteria can be divided into species that exert or potentially harmful or beneficial effects on the host. Pathogenic effects (which may be caused by clostridia or bacteroids, for example) include diarrhea, infections, liver damage, carcinogenesis and intestinal putrefaction. The health-promoting effects can be caused by inhibiting the growth of harmful bacteria, stimulating immune functions, improving digestion and absorbing essential nutrients, and synthesizing vitamins. An increase in numbers and / or activities of bacterial groups (such as Bifidobacterium and Lactobacillus) that may have health-promoting properties is desired.

Specifically for infants just before birth, a baby's gastrointestinal tract is thought to be sterile. During the process of birth, it finds bacteria from the digestive tract and the mother's skin and begins to be colonized. There are large differences in the composition of the intestinal microflora in response to infant feeding. The fecal flora of breastfed infants include appreciable populations of Bifidobacteria with some species of Lactobacillus, while infants fed formula have more complex microflora, with Bifidobacteria, Bacteroides, Clostridia and Streptococci all normally present. After weaning, a pattern of intestinal microflora that resembles the pattern of adults is established. Breast milk is recommended for all infants. However, in some cases, breastfeeding is unsuitable or unsuccessful for medical reasons or the mother chooses not to breastfeed. Infant formulas have been developed for these situations.

One approach to promoting the numbers and / or activities of beneficial bacteria in the colon is the addition of prebiotics to food products. A prebiotic is an indigestible food ingredient that advantageously affects the host by selectively stimulating the growth and / or activity of one or a limited number of colon bacteria and thus improves the health of the host. These ingredients are non-digestible in the sense that they are not degraded and absorbed in the stomach or small intestine and therefore pass intact in the colon where they are selectively fermented by the beneficial bacteria. Examples of prebiotics include certain oligosaccharides, such as fructooligosaccharides (FOS) and galactooligosaccharides (GOS).

It is known that human milk contains a greater amount of indigestible oligosaccharides than most other animals' milks. In fact, indigestible oligosaccharides represent the third largest solid component (after lactose and lipids) in breast milk, occurring at a concentration of 12-15 g / L in colostrum and 5-8 g / L in mature milk. Oligosaccharides from human milk are very resistant to enzymatic hydrolysis, indicating that these oligosaccharides may have essential functions not directly related to their calorific value. As the composition of human milk is better understood, the addition of prebiotics to the infant formula has also been proposed. A number of infant formulas supplemented with prebiotics, such as mixtures of fructo-oligosaccharides and galacto-oligosaccharides, for example, are available commercially. However, such mixtures only roughly approach the oligosaccharide mixture of human milk. More than 100 different oligosaccharide components were detected in human milk, some of which have so far not been detected in animal milks, such as cow's milk, or have only been detected in small amounts. Examples of classes of human milk oligosaccharides which are present in cow's milk and colostrum only in very small amounts or are not present are sialylated and fucosylated oligosaccharides. US Patent Application No. 2003/0129278 describes a mixture of oligosaccharides based on oligosaccharides produced from one or more animal milks, which is characterized in that it comprises at least two fractions of oligosaccharides each consisting of, at least two different oligosaccharides, which do not include free lactose. The total spectrum of the oligosaccharides present in the oligosaccharide mixture differs from those present in animal milk or animal milks from which the oligosaccharide fractions have been extracted. In addition a) if said oligosaccharides are extracted from a single animal milk, the ratio of neutral oligosaccharides to acid (sialylated) oligosaccharides is 90-60: 10-40% by weight, or b) if said oligosaccharides are extracted from, at least two animal milks, the oligosaccharides extracted from two different animal milks constitute each 10% by weight of the total amount of oligosaccharides present in the oligosaccharide mixture.

It is an aim of the invention to provide a mixture of oligosaccharides which is effective as a prebiotic, particularly in the human gut.

Summary of the Invention

In one aspect, the invention relates to a mixture of oligosaccharides comprising 5-70% by weight of at least one N-acetylated oligosaccharide selected from the group comprising GalNAcal, 3Gaipi, 4Glc and Gaipi, 6GalNAcOCl, 3Gaipi, 4Glc, 20 -90% by weight of at least one neutral oligosaccharide selected from the group comprising GalNAcCCl, 3Gaipi, 4Glc and Gaipi, 6GalNAcCCl, 3Gaipi, 4Glc, 20-90% by weight of at least one neutral oligosaccharide selected from the group comprising Galpl, 6Gal, Galpl, 6Galpl, 4Glc Galpl, 6Galpl, 6Glc, Gal3l, 3Gal3l, 3Glc, GaiPl, 3Gaipi, 4Glc, Galpl, Galpl, 6Galpl, 4Glc, Gaipi, 6Gal3l, 3Gal3l, 4Glc Gaipi, 3Gaipi, 6Gaipi, 4Glc and Gaipi, 3Gaipi, 3Gaipi, 4Glc and 5-50% by weight of at least one sialylated oligosaccharide selected from the group comprising NeuAcd2,3Gaipi, 4Glc and NeuAca2,6Gaipi, 4Glc.

This ingredient is a novel protective and immunomodulatory ingredient which is particularly effective as a prebiotic. The blend is structurally closer to the oligosaccharides of human breast milk than commercially available prebiotic ingredients, such as GOS and FOS, for example, in that it includes a mixture of acidic and neutral oligosaccharides.

In one embodiment, the oligosaccharide mixture may be derived from animal milk, such as one or more of cow's milk, goat's milk or buffalo's milk.

In another aspect, the invention relates to a food product comprising a mixture of oligosaccharides, as described above. Optionally, the food product is a food or formula for infants, but the product may be any food or beverage consumed by infants, children or adults. Consumption of a food product containing one of these mixtures of oligosaccharides as a prebiotic will selectively promote the growth and / or activity of one or a limited number of beneficial bacteria in the colon and thus improve the health of the host.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows counts of Bifidobacterium breve, (A, B) B. longum (C, D) and C. perfringens (E, F) in the small intestine (jejunum) and feces after 2 weeks of the intervention described in Example 3. Mean bacterial counts expressed as log values with robust standard deviation are plotted for the control group and for the OS blend group according to the invention. The probabilities of an effect are indicated on the basis of robust ANOVA. (N = 9 to 10). (NA, not applicable due to values below the limit of detection). Figure 2 shows the relative overall metabolic activity of C. perfringens during the feeding time of the intervention described in Example 3. The data is represented as a &quot; &quot; whisker box diagram &quot; with the median and interquartile range of the control group (gray boxes) and the group with the OS blend (white boxes). (N = 9 to 10). The prebiotic effect was significant for day 0 and day 14, as assessed by the median test involving Fisher's Exacto test (p <0.005). Figure 3 shows counts of Bifidobacterium breve, B. longum and C. perfringens in the small intestine (jejunum) (A, C, E) and feces (B, D, F) after 2 weeks of intervention. Medians of the bacterial counts expressed as log values with interquartile ranges are plotted for the control group, the group only with neutral galacto-oligosaccharides and the group with the oligosaccharide mixture according to the invention. 6 ND, not detected.

Detailed description of the invention

In the present description, the following words have a definition which must be taken into account in the reading and interpretation of the description, examples and claims. &quot; Infant Formula &quot;: Food product intended for the complete nutrition of infants during the first four to six months of life. (Article 1.2 of the European Commission Directive 91/321 / EEC of 19 May 1991 on infant formulas and follow-on formulas).

It has to be understood that infants can be fed exclusively with infant formulas or that an infant formula can be fed by the mother or other care provider as a complement to human milk. The term &quot; infant formula &quot; as used herein is synonymous with the commonly used &quot; starter formula &quot;

N-acetylated oligosaccharides: oligosaccharides having an N-acetyl residue.

Neutral oligosaccharides: the oligosaccharides which have no charge and no N-acetyl residue. &quot; Prebiotic &quot;: an nondigestible food ingredient that beneficially affects the host by selectively stimulating the growth and activity of one or a limited number of colon bacteria and thus enhances the health of the host. (Gibson and Roberfroid, &quot; Dietary Modulation of the Human Colonic Microbiota: Introducing the Concept of Prebiotics &quot;, J. Nutr. 125: 1401-1412). &quot; Oligosaccharide &quot;: carbohydrate having a degree of polymerization (DP) ranging from 2 to 20, inclusive, but not including lactose.

Sialylated oligosaccharides: oligosaccharides having a sialic acid residue with associated charge. The invention provides a mixture of oligosaccharides comprising 5-70% by weight of at least one N-acetylated oligosaccharide selected from the group comprising GalNAcocl, 3Gaipi, 4Glc and Gaipi, 6GalNAcCCl, 3Gaipi, 4Glc, 20-90% by weight of at least one neutral oligosaccharide selected from the group comprising Gaipi, 6Gal, Gaipi, 6Gaipi, 4Glc Galpl, 6Galpl, 6Glc, Gaipi, 3Gaipi, 3Glc, Gaipi, 3Gaipi, 4Glc, Gaipi, 6Galpl, 6GaI, 4Glc, Galpl, Gaipi, 3Gaipi, 4Glc Galpl, 3Galpl, 6Galpl, 4Glc and Gaipi, 3Gaipi, 3Gaipi, 4Glc and 5-50% by weight of at least one sialylated oligosaccharide selected from the group comprising NeuAca2,3Gaipi, 4Glc and NeuAca2,6Gaipi, 4Glc and infant or adult food products comprising such an oligosaccharide mixture.

Preferably, the blend comprises 10-70% by weight of the specified N-acetylated oligosaccharide (s), 20-80% by weight of the specified neutral oligosaccharide (s) and 10-50% by weight of the sialylated oligosaccharide (s) specified. More preferably, the blend comprises 15-40% by weight of the N-acetylated oligosaccharide (s), 40-60% by weight of the other neutral oligosaccharide (s) and 15-30% by weight of the sialylated oligosaccharides. A particularly preferred blend is 30% by weight of the N-acetylated oligosaccharide (s), 50% by weight of the neutral oligosaccharide (s) and 20% by weight of the sialylated oligosaccharide (s).

Alternatively, the blend may advantageously comprise 5-20% by weight of the specified N-acetylated oligosaccharide (s), 60-90% by weight of the specified neutral oligosaccharides and 5-30% by weight of the specified sialylated oligosaccharides. The oligosaccharide mixture of the invention may be prepared from one or more animal milk. Milk can be obtained from any animal, particularly cows, goats, buffaloes, horses, elephants, camels or sheep.

Alternatively, the oligosaccharide mixture may be prepared by acquisition and blending of the individual components. For example, galacto-oligosaccharides synthesized, such as Galpl, 6Galpl, 4Glc Galpl, 6Glc, 6Glc, Galpl, 3Galpl, 4Glc, Galpl, 6Galpl, 6GaI, 4Glc, Galpl, 6Galpl, 3Galpl, 4Glc and Gaipi, 3Gaipi , â € ƒâ € ƒâ € ƒGaipil, 4Glc and mixtures thereof are commercially available under the tradenames Vivinal® and Elix®®. Other suppliers of oligosaccharides are Dextra Laboratories, Sigma-Aldrich Chemie GmbH and Kyowa Hakko Kogyo Co., Ltd. Alternatively, specific glycosyltransferases such as galactosyltransferases can be used to produce neutral oligosaccharides.

The N-acetylated oligosaccharides can be prepared by the action of glucosaminidase and / or galactosaminidase N-acetyl glucose and / or N-acetylgalactose. Likewise, N-acetyl-galactosyl transferases and / or N-acetyl-glycosyl transferases may be used for this purpose. The N-acetylated oligosaccharides can also be produced by fermentation technology using the respective enzymes (recombinant or natural) and / or microbial fermentation. In the latter case, the microorganisms may express their natural enzymes and substrates or they may be genetically engineered to produce the respective substrates and enzymes. Single microbial cultures or mixed cultures may be used. The formation of N-acetylated oligosaccharides can be initiated by acceptor substrates starting from any degree of polymerization (DP) of DP = 1 onwards. Another option is the chemical conversion of ketohexoses (e.g., fructose) free or bound to an oligosaccharide (e.g., lactulose) to N-acetylhexosamine or an oligosaccharide containing N-acetylhexosamine as described in Wrodnigg, T.M.; Stutz, A. E. (1999) Angew. Chem. Int. Ed. 38: 827-828.

The sialylated oligosaccharides 3'-sialyl-lactose and 6'-sialyl-lactose can be isolated by chromatographic or filtration technology from a natural source, such as animal milks. Alternatively, they may also be produced by biotechnology using specific sialyltransferases by enzyme-based fermentation technology (recombinant or natural) or by microbial fermentation technology. In the latter case, the microorganisms may express their natural enzymes and substrates or they may be genetically engineered to produce the respective substrates and enzymes. Individual microbial cultures or mixed cultures may be used. The formation of sialyl-oligosaccharides can be initiated by acceptor substrates from any degree of polymerization (DP) of DP = 1 onwards.

In a preferred aspect of the invention, the oligosaccharide mixtures described above are incorporated into a food product. In the context of the present invention, the term &quot; food product &quot; intended to cover any consumable matter. Thus, it may be a product intended for human consumption, in particular infant formula, transition formula, baby food such as baby cereal and the like. In particular, the oligosaccharide mixture of the invention may be incorporated into infant formulas, dehydrated milk or cereal mixtures. The food product may be prepared by any suitable mode known in the art and according to the type of product and the oligosaccharide mixture of the invention may be added to the product in an appropriate step of the manufacturing process. For example, an infant formula may be prepared by mixing the protein source, any non-lactose carbohydrate, and the source of fat in appropriate proportions. Emulsifiers may be added if desired. At this point vitamins and minerals can be added but are usually added later to prevent thermal degradation. Any lipophilic vitamins, emulsifiers and the like may be dissolved in the source of fat prior to mixing. Water, preferably water which has been subjected to reverse osmosis, may then be mixed to form a liquid mixture. The liquid mixture can then be thermally treated to reduce bacterial load. For example, the liquid mixture may be rapidly heated to a temperature in the range of about 80 ° C to about 110 ° C for about 5 seconds to about 5 minutes. This can be accomplished by steam injection or by heat exchange, for example in a plate heat exchanger. The liquid mixture can then be cooled to about 60øC to about 85øC, for example by rapid cooling. The liquid mixture can then be homogenized, for example, in two steps at about 7 MPa to about 40 MPa in the first step and from about 2 MPa to about 14 MPa in the second step. The homogenized blend may then be further cooled to the addition of heat-sensitive components, such as vitamins and minerals. The pH and solids content of the homogenized mixture are conveniently normalized at this point. The homogenized mixture is transferred to a suitable drying apparatus, such as a spray dryer or lyophilizer, and converted into powder. The powder should have a moisture content of less than about 5% by weight. The oligosaccharide mixture of the invention is preferably directly added to the formula for infants by dry blending. However, if it has been prepared from animal milk, for example as described below, it may be convenient to add the oligosaccharide mixture without first removing all the lactose. As the infant formula contains a carbohydrate component which is often wholly or partly comprised of lactose, it will be apparent to one skilled in the art that the amount of carbohydrate in the infant formula will need to be adjusted to take into account the carbohydrates which will be provided by the oligosaccharide mixture. The final concentration of the oligosaccharide mixture in the infant food product or infant formula is preferably from 0.3 to 4%, preferably from 0.75 to 1.54% by weight of dry matter. This corresponds to a concentration of 0.2 to 5 grams per liter of reconstituted formula, preferably 1 to 2 g / L. However, these amounts should not be considered as limiting and should be adapted to the target population, for example based on the weight and age or health of the infant or infant. Preferably, the formulation or feed containing the oligosaccharide mixture of the invention is given to the baby at each meal.

Alternatively, the oligosaccharide mixture may be added to wet or infant food products by wet mixing. The blend may be added to the formula for infants or infants in concentrations of about 0.2 to 5 grams of oligosaccharides per liter of product. However, these amounts should not be considered as limiting and should be adapted to the target population, for example based on the weight and age of the infant or infant, or the health of the specific population.

While it is preferred to supplement food products specifically targeted to nutrition for infants or infants, it may be advantageous to supplement food products not specifically targeted, or targeted to the adult population. For example, the oligosaccharide mixtures of the invention may be incorporated into dietary nutritional products and nutritional products for the elderly. These food products may include milk, yogurt, curd cheese, cheese, fermented milk, fermented milk products, ice creams, fermented cereal products or milk-based products, among others.

In addition to the oligosaccharide mixture of the invention, a food product, such as an infant formula, may include one or more additional oligosaccharides which are added separately. The invention will now be illustrated by reference to the following examples.

Example 1

A method of preparing a mixture of oligosaccharides according to the invention will now be described by way of example only. 200,000 liters of a whey ultrafiltration permeate are preconcentrated to 22% (w / w) total solids (TS), pasteurized at about 75 ° C for about 30 seconds and then concentrated by evaporation to 60 ° C to achieve a TS of 59% (w / w). The liquid is cooled in a crystallizer at a rate of 2 ° C per hour for a period of 24 hours to crystallize the lactose. The crystallized lactose is washed and then withdrawn by a juicer. The remaining liquid (mother liquor) is clarified through a decanter. The 77000 liters with 17.7% TS obtained from the clarifier are reconcentrated by evaporation at 60 ° C to give a TS of 55% (w / w) and subjected to a second step of crystallization of lactose under the same conditions as above. The 29,000 liters at 20.55 TS so obtained are demineralized by a combination of electrodialysis and ion exchange in a manner known per se to yield 28500 liters of a 90% demineralized liquor with 17.3% TS. This liquid, which contains approximately 1.5 grams per liter of a blend of about 30% by weight of GalNAcal, 3Gaipi, 4Glc and GaiPl, 6GalNAcOCl, 3Gal3l, 4Glc, 50% by weight of Gaipi, 6Gaipi, 6Glc, Gaipi, 6Gaipi, 4Glc and Gaipi, 3Gaipi, 4Glc and 20% by weight NeuAca2,3Gaipi, 4Glc and NeuAca2,6Gaipi, 4Glc, depending on the starting material, can be added directly to a food product, such as an infant formula or can be further concentrated in a manner known per se by those skilled in the art.

For example, the lactose remaining in the solution can be hydrolyzed to glucose and galactose and these monosaccharides can be removed by nanofiltration or, if desired, the galactose can be at least partially polymerized. for example, by the action of β-galactosidase to produce galacto-oligosaccharides which will also be retained by the nanofiltration membrane.

An example of the composition of an infant formula containing a preparation according to the present invention is presented below.

Nutrient per 100 kcal per liter Energy (kcal) 100 670 Protein (g) 1.83 12.3 Fats (g) 5.3 35.7 Linoleic acid (g) 0.79 5.3 (continued)

Nutrient per 100 kcal per liter α-linolenic acid (mg) 101 675 Lactose (g) 11.2 74.7 OS Mixture of Example 1 (g) 0.15 1.0 Minerals (g) 0.37 2.5 Na (mg) 23 150 K (mg) 89 590 Cl (mg) 64 430 Ca (mg) 62 410 P (mg) 31 210 Mg (mg) 7 50 Mn (pg) 8 50 Se (pg) 2 13 Vitamin A (pg RE) 105 700 Vitamin D (pg) 1.5 10 Vitamin E ((pg TE) 0.8 5.4 Vitamin Kl (pg) 8 54 Vitamin C (mg) 07 0.47 Vitamin B2 (mg) 0, 15 1.0 Niacin (mg) 1 6.7 Vitamin B6 (mg) 0.075 0.50 Folic acid (pg) 9 60 Pantothenic acid (mg) 0.45 1 Vitamin B12 (pg) 0.3 2 Biotin (pg) 2.2 15 Choline (mg) 10 67 Fe (mg) 1.2 8 (pg) 15 100 16 (continued)

Nutrient per 100 kcal per liter Cu (mg) 0.06 0.4 Zn (mg) 0.75 5

The effect of a mixture of oligosaccharides according to the invention on the establishment and composition of the intestinal microflora was investigated in mice free of microorganisms.

C3H mice were kept free of microorganisms up to the age of 8 weeks and fed on a semisynthetic AIN diet. On day -1 of the intervention, a single dose of a human baby microflora cocktail was administered to each mouse by catheter. The mice were divided into 2 groups and their diet was changed to an AIN diet containing 1.2% (w / w) lactose as additional carbohydrate and to the other group 1.2% (w / w) ) of lactose and 2.5% (w / w) of a mixture of oligosaccharides according to the invention comprised of 5% (w / w) sialyl-oligosaccharides, 5% (w / w) N-acetylated oligosaccharides and 90% (w / w) neutral oligosaccharides. The microflora establishment was evaluated in the small intestine and feces after 14 days of intervention by counting Bifidobacterium breve, B. longum and Clostridium perfringens plaques. From Figure 1 it can be seen that both resident bifidobacteria showed increased counts in the small intestine and especially in the faeces. On the other hand, the counts of C. perfringens were reduced in the small intestine and especially in the feces.

Over the feeding time, the relative metabolic activity of C. perfringens was monitored by determination of the 16S RNA levels. Briefly, the RNA was extracted from freshly collected fecal samples and the RNA was subjected to an RT-PCR reaction to amplify 16S rRNA. The PCR products were separated by denaturing gradient gel electrophoresis and the 16S rRNA of C. perfringens was quantified and normalized to the signal of 16S E. coli rRNA which remained constant during the feeding time. As can be seen in Figure 2, only one day after the intervention (Day 0 in Figure 2) is that the metabolic activity of C. perfringens was considerably and significantly reduced by the OS blend according to the invention, compared to the group and remained the lowest during the duration of the intervention.

Example 4 The effect of a mixture of oligosaccharides according to the invention on the establishment and composition of the intestinal microflora was compared with the effect of neutral oligosaccharides alone on gnotobiotic mice.

C3H mice were kept free of microorganisms up to the age of six weeks and fed with a semi-synthetic AIN diet. A single dose of a human baby microflora cocktail was administered to each mouse by gavage and the microflora was allowed to settle for two weeks. Mice were divided into 3 groups and their diet was changed to an AIN-containing diet containing the additional 1, 2% (w / w) lactose group as the additional carbohydrate 1.2% (w / w) for the second group 1.2% w / w) of lactose and 2.5% (w / w) galactooligosaccharides and for the third group 1.2% (w / w) lactose and 2.5% (w / w) of a mixture of oligosaccharides according to the invention comprised of 5% (w / w) sialyl-oligosaccharides, 5% (w / w) N-acetylated oligosaccharides and 90% (w / w) neutral oligosaccharides. The establishment of the microflora was evaluated in the small intestine and feces after 14 days of intervention by counting Bifidobacterium breve, B. longum and Clostridium perfringens plaques. From Figure 3 it can be seen that both resident bifidobacteria showed increased counts in the small intestine and especially in faeces in the presence of the OS mixture in the diet. In the presence of only neutral galacto-oligosaccharides, the resident B. longum did not present increased counts in both the small intestine and faeces. The counts of C. perfringens were reduced in the small intestine and also in the faeces in the presence of the OS mixture according to the invention. However, in the presence of only neutral galacto-oligosaccharides, increased levels of C. perfringens in feces were observed. Taken together, these results strongly suggest that the oligosaccharide mixture according to the invention has equilibrium effects of the microbiota which are superior to those of the neutral oligosaccharides alone.

Lisbon, June 14, 2010 19

Claims (7)

A mixture of oligosaccharides comprising 5-70% by weight of at least one N-acetylated oligosaccharide selected from the group comprising GalNAcCCl, 3Gaipi, 4Glc and Gaipi, 6GalNAcal, 3Gaipi, 4Glc, 20-90% by weight of, at least one neutral oligosaccharide selected from the group comprising Gaipi, 6Gal, Galpl, 4Glc Galpl, 6Glpl, 6Glc, Galpl, 3Gpl, 3Glc, Gaipi, 3Gaipi, 4Glc, Gal3l, 6Gal3l, 6Gal3l, 4Glc, Galpl, δGalpl, 3Galpl , 4Glc Gaipi, 3Gaipi, Galipi, 4Glc and Galpl, 3Galpl, 3Galpl, 4Glc and 5-50% by weight of the at least one sialylated oligosaccharide selected from the group comprising NeuAc (X2,3Gaipi, 4Glc and NeuAca2,6Ga11, 4Glc. A mixture of oligosaccharides as claimed in Claim 1 which comprises 10-70% by weight of the N-acetylated oligosaccharides, 20-80% by weight of the neutral oligosaccharides and 10-50% by weight of the sialylated oligosaccharides. A mixture of oligosaccharides as claimed in Claim 1 or 2 which comprises 15-40% by weight of the N-acetylated oligosaccharides, 40-60% by weight of the neutral oligosaccharides and 15-30% by weight of the sialylated oligosaccharides. 1 A mixture of oligosaccharides as claimed in any preceding claim comprising 30% by weight of the N-acetylated oligosaccharides, 50% by weight of the neutral oligosaccharides and 20% by weight of the sialylated oligosaccharides. A mixture of oligosaccharides as claimed in Claim 1 comprising 5-20% by weight of the N-acetylated oligosaccharides, 60-90% by weight of the neutral oligosaccharides and 5-30% by weight of the sialylated oligosaccharides. A mixture of oligosaccharides as claimed in any preceding claim which is derived from animal milk. A mixture of oligosaccharides as claimed in Claim 6 which is derived from one or more of cow's milk, goat's milk or buffalo's milk. A mixture of oligosaccharides as claimed in any of claims 1 to 6 which is made synthetically. A food product comprising a mixture of oligosaccharides as claimed in any preceding claim. A food product as claimed in Claim 9 which is an infant formula. A food product as claimed in Claim 9 or 10 which comprises 0.3 to 4% by weight based on dry matter of the oligosaccharide mixture. Lisbon, June 14, 2010 3 1/3 Figure 1. In the small intestine In the feces 5 o &gt; &gt; 4 log (æg / g λ of tissue -Q 3 CQ 2 ε D 7 6 O) 5 æ log (æg / g Q of tissue ½ O 2 wc 3.5 0) D 5 3.0 C tissue 2.5%) Q. 2.0. ò A10 I. B p (a) = 0.0000 C 1110 lag (ufe / g tissue 7 6 5 D 7.0 6.5 log (ufo / g of fabric 6-0 Control Mixture of OS 5.5 5.0 Figure 2 Day 14 Day 0 100 80 60 Relative metabolic activity (% of reference) 40 20 Day 0 Day 1 Control Mixture of Control OS Mixture Control Mixture of OS 0 3/3 Figure 3. In slack gut In feces Neutral OS Control OS Mix Control Neutral OS Mix of the